Surface modulation of extracellular vesicles with cell-penetrating peptide-conjugated lipids for improvement of intracellular delivery to endothelial cells.
Surface modulation of extracellular vesicles with cell-penetrating peptide-conjugated lipids for improvement of intracellular delivery to endothelial cells.
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DOI：
10.1016/j.reth.2022.12.007
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发表时间：
2023-03
影响因子：
4.3
通讯作者：
Teramura, Yuji
中科院分区：
文献类型：
--
作者：
Huang, Tianwei;Sato, Yuya;Kuramochi, Akiko;Ohba, Yoshio;Sano, Masayuki;Miyagishi, Makoto;Tateno, Hiroaki;Wadhwa, Renu;Kawasaki, Kazunori;Uchida, Takeyuki;Ekdahl, Kristina N.;Nilsson, Bo;Chung, Ung-il;Teramura, Yuji
关键词：
外泌体（直径30-200 nm）是由含有DNA、微小RNA（miRNA）和蛋白质的细胞分泌的细胞外囊泡的亚型。预期外泌体作为在疾病治疗中递送药物或功能性miRNA的手段是有价值的。然而，外泌体的递送并不足够有效，即使外泌体具有内在递送功能。细胞穿透肽（CPP）是促进细胞摄取分子和囊泡的短肽家族。我们以前报道过，修饰的细胞和脂质体与CPP共轭的脂质，共轭与聚（乙二醇）共轭磷脂（PEG-脂质），诱导粘附的CPP，可以用于基于细胞的测定和收获脂质体。在这项研究中，我们的目的是使用达特肽（YGRKKRRQRRR）-PEG-脂质来调节外泌体表面，以改善向内皮细胞的细胞内递送。我们从HEK 293 T细胞培养物的培养基中分离并表征了外泌体。使用异硫氰酸荧光素标记将具有不同间隔分子量和脂质类型的达特缀合的PEG-脂质掺入外来体中，以优化固定在外泌体表面上的Tat-PEG-脂质的数量。将用Tat-PEG-脂质修饰的外泌体与人脐静脉内皮细胞（HUVEC）孵育以研究相互作用。通过流式细胞术在HUVEC内高度检测到与掺入在外泌体表面上的5 kDa PEG和C16脂质缀合的达特。对于对照组，HUVEC中的荧光可忽略不计。因此，可以在外泌体表面上修饰Tat-PEG-脂质，从而改善外泌体的细胞内递送。
Exosomes (diameter 30–200 nm) are a subtype of extracellular vesicles secreted by cells containing DNA, microRNA (miRNA), and proteins. Exosomes are expected to be valuable as a means of delivering drugs or functional miRNAs in treatment of diseases. However, the delivery of exosomes is not sufficiently effective, even though exosomes have intrinsic delivery functions. Cell-penetrating peptides (CPPs) are short peptide families that facilitate cellular intake of molecules and vesicles. We previously reported that the modification of cells, and liposomes with CPP-conjugated-lipids, CPPs conjugated with poly (ethylene glycol)-conjugated phospholipids (PEG-lipid), that induce adhesion by CPPs, can be useful for cell-based assays and harvesting liposomes. In this study, we aimed to modulate the exosome surface using Tat peptide (YGRKKRRQRRR)-PEG-lipids to improve intracellular delivery to endothelial cells. We isolated and characterized exosomes from the medium of HEK 293 T cell cultures. Tat conjugated PEG-lipids with different spacer molecular weights and lipid types were incorporated into exosomes using fluorescein isothiocyanate labeling to optimize the number of Tat-PEG-lipids immobilized on the exosome surface. The exosomes modified with Tat-PEG-lipids were incubated with human umbilical vein endothelial cells (HUVECs) to study the interaction. Tat conjugated with 5 kDa PEG and C16 lipids incorporated on the exosome surface were highly detected inside HUVECs by flow cytometry. Fluorescence was negligible in HUVECs for control groups. Thus, Tat-PEG-lipids can be modified on the exosome surface, improving the intracellular delivery of exosomes.
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DOI：
10.1002/prca.201200134
发表时间：
2013-10
期刊：
Proteomics. Clinical applications
影响因子：
--
作者：
Nyalwidhe JO;Betesh LR;Powers TW;Jones EE;White KY;Burch TC;Brooks J;Watson MT;Lance RS;Troyer DA;Semmes OJ;Mehta A;Drake RR
通讯作者：
Drake RR
影响因子：
9.5
作者：
Teramura, Yuji;Asif, Sana;Nilsson, Bo
通讯作者：
Nilsson, Bo
影响因子：
10.2
作者：
Wang, Congcong;Li, Ning;Li, Yutian;Hou, Shasha;Zhang, Wenxin;Meng, Zhaowei;Wang, Shen;Jia, Qiang;Tan, Jian;Wang, Renfei;Zhang, Ruiguo
通讯作者：
Zhang, Ruiguo
源自脓毒症小鼠血清的外泌体通过外泌体相关细胞因子调节免疫反应
影响因子：
7.3
作者：
Gao, Kun;Jin, Jingmiao;Jiang, Yong
通讯作者：
Jiang, Yong
影响因子：
4.6
作者：
Ohgaki R;Teramura Y;Hayashi D;Quan L;Okuda S;Nagamori S;Takai M;Kanai Y
通讯作者：
Kanai Y